Objetivos: Verificar a frequência de Chlamydia trachomatis e a sua associação com a expressão de p16/Ki-67 em mulheres com lesões intraepiteliais cervicais. Analisar a associação entre a positividade para Chlamydia trachomatis e variáveis demográficas selecionadas, antecedentes sexuais e obstétricos, resultados anormais citológicos e anatomopatológicos. Também verificar a associação entre a expressão da dupla coloração para p16/Ki-67 com os resultados citológicos e anatomopatológicos. Métodos: Estudo de corte transversal em 1481 mulheres de 18 a 64 anos, participantes de projeto de rastreamento para câncer cervical realizado em São Paulo. As citologias foram coletadas em meio líquido e no líquido residual foi pesquisada a presença do DNA da Chlamydia trachomatis. Nos resultados de citologia anormal foi feita a pesquisa da expressão das proteínas p16/Ki-67. A análise estatística foi realizada usando-se os testes qui-quadrado e da razão de verossimilhanças. Foram estimados os valores de odds ratios (OR) com os respectivos intervalos com 95% de confiança. Resultados: A frequência de detecção da Chlamydia trachomatis foi 15,6%. Não houve associação estatisticamente significativa entre a presença de Chlamydia trachomatis e a expressão de p16/Ki-67 [OR=1,35 (0,5-3,4)]. A idade e o número de parceiros sexuais apresentou associação significativa com a presença de Chlamydia trachomatis [OR= 2,01 (1,1-3,6) e 4,14 (1,7-10,3)]. Não foi encontrada associação significativa entre citologia alterada e a positividade para Chlamydia trachomatis [1,21 (0,46-3,2)]. Não foi observada associação significativa entre os resultados anatomopatológicos e a presença de Chlamydia trachomatis (p = 0,112). A expressão do p16/Ki-67 mostrou associação estatisticamente significativa com lesões intraepiteliais cervicais de alto grau tanto nos resultados citológicos quanto anatomopatológicos. Conclusões: A frequência de infecção por Chlamydia trachomatis na amostra estudada foi de 15,6%. A associação da Chlamydia trachomatis com a dupla coloração para p16/Ki-67 nas citologias anormais não foi significativa, não sendo possível estabelecer uma associação clara entre a presença de Chlamydia trachomatis e a persistência da infecção por HPV oncogênico detectada por este marcador. Dentre as variáveis demográficas pesquisadas, a faixa etária apresentou associação estatisticamente significativa com a presença do DNA da Chlamydia trachomatis; mulheres com idade entre 35 e 45 apresentaram a maior frequência da infecção; entretanto, mesmo as outras faixas etárias mostraram uma alta frequência da presença desse patógeno; foi observada maior frequência da infecção entre as mulheres com mais de 10 parceiros sexuais, quando comparadas com as com menor número de parceiros durante a vida e essa associação foi estatisticamente significativa; não foi demonstrada associação significativa entre os resultados anormais da citologia com a positividade para a presença do DNA da Chlamydia trachomatis. Não foi encontrada associação significativa entre os resultados anatomopatológicos dirigidas pela colposcopia com a positividade da infecção pela bactéria. A positividade da dupla coloração para p16/Ki-67 foi significativamente maior nas lesões intraepiteliais cervicais de alto grau. Foi demonstrada associação estatisticamente significativa entre a expressão do p16/Ki-67 com os resultados anatomopatológicos das biopsias dirigidas pela colposcopia / Objective: To verify the frequency of Chlamydia trachomatis and its association with the expression of p16/Ki-67 on women with cervical intraepithelial lesions. To analyze the association between Chlamydia trachomatis presence and the selected demographic variables such as sexual and obstetric history, abnormal cytology and histopathology, as well as analyzing any association between the expression of dual staining for p16/Ki-67 with cytological and histopathological results. Methods: Cross-sectional study on 1481 women with ages between 18 and 64 years, who were enrolled in a screening project for cervical cancer held in São Paulo. The cytology was collected in liquid based medium and the residual liquid was submitted for examination to find the presence of Chlamydia trachomatis DNA. The expression of protein p16/Ki-67 was performed in the abnormal cytology results. Statistical analysis was performed using chi-square tests and likelihood ratio. The values of the odds ratios (OR) with respective intervals of 95% confidence were estimated. Results: The frequency of detection of Chlamydia trachomatis was 15.6%. There was no statistical significant association between the presence of Chlamydia trachomatis and the expression of p16/Ki-67 [OR = 1.35 (0.5 to 3.4)]. Both the age and number of sexual partners presented a significant association in presence of Chlamydia trachomatis [OR = 2.01 (1.1 to 3.6) and 4.14 (1.7 to 10.3)]. There was no significant association between abnormal cytology and positivity for Chlamydia trachomatis [1.21 (0.46 to 3.2)]. No significant association was found between histopathological results and presence of Chlamydia trachomatis (p= 0.112). The expression of p16/Ki-67 showed a significant statistical association with high grade intraepithelial lesions in both cytological and histopathological results. Conclusions: The prevalence of Chlamydia trachomatis infection on the sample studied was 15.6%. The association of Chlamydia trachomatis with dual staining for p16/Ki-67 in abnormal cytology was not significant, therefore, it is not possible to establish a clear association between the presence of Chlamydia trachomatis and the persistence of oncogenic HPV infection detected by this marker. Among the demographic variables, the age range showed statistically significant association with the presence of Chlamydia trachomatis; women aged between 35 - 45 years showed the highest rate of infection, nevertheless the other age ranges showed a high frequency of the presence of this pathogen; It has been observed a higher number of infected women who had more than 10 sexual partners compared to woman who had less partners throughout life and this association was statistically significant; No significant association was found between abnormal cytology with positivity for the presence of Chlamydia trachomatis DNA; There was no significant association between pathological results directed by colposcopy with the positivity of Chlamydia trachomatis; The positivity of the double staining for p16/Ki-67 was significantly higher in the cervical high-grade intraepithelial lesions; Statistical significant association was demonstrated between the expression of p16/Ki-67 with histopathological results of biopsy directed by colposcopy

Orientadores: Luiz Carlos Zeferino, Silvia Helena Rabelo dos Santos. / Tese (doutorado) - Universidade Estadual de Campinas, Faculdade de Ciências Médicas / Made available in DSpace on 2018-08-21T17:32:08Z (GMT). No. of bitstreams: 1 CastroSobrinho_JucaraMariade_D.pdf: 1214452 bytes, checksum: db87fc4f5e35e2d9e1091af5b48fc8cf (MD5) Previous issue date: 2012 / Resumo: Objetivo: Analisar a associação entre a coinfecção por papilomavírus humano (HPV), Chlamydia trachomatis (CT), vaginose bacteriana (VB) e resposta inflamatória (RI) com a gravidade da neoplasia cervical. Sujeitos e métodos: Estudo experimental, de corte transversal, realizado em Campinas, São Paulo e em Goiânia, Goiás, Brasil. A casuística incluiu amostras biológicas de 290 mulheres consecutivas submetidas à excisão da zona de transformação (EZT) ou conização. Para o estudo que avaliou a coinfecção entre HPV e CT e a associação com gravidade da neoplasia cervical foram selecionadas 251 (86,6%) mulheres infectadas por HPV de alto risco (HR-HPV). A detecção de HPV foi realizada utilizando os primers PGMY09/11 e a genotipagem através de hibridização reversa em pontos. A detecção da CT foi realizada por polimerase chain reaction (PCR) empregando primers cujo alvo é uma região de plasmídio críptico, gerando um fragmento de aproximadamente 512 pares de base. Para o estudo que avaliou a VB e resposta inflamatória e a associação destas condições com a gravidade da neoplasia cervical foram selecionadas 211 mulheres infectadas por HR-HPV, com esfregaços cervicais disponíveis para as análises. A presença de 20% ou mais de células indicadoras no esfregaço cervical corado pelo método de Papanicolau foi considerada positiva para VB. A resposta inflamatória nos esfregaços cervicais foi avaliada pela contagem do número de neutrófilos. O encontro de 30 ou mais neutrófilos por campo microscópico, observado sob o aumento de 1000x, foi considerado como presença de resposta inflamatória. Resultados: A prevalência de CT em mulheres HPV positivas foi de 15,1% (38/251). Foi observada uma associação significativa em mulheres CT negativas, com 30 anos ou mais, e NIC 2 ou pior diagnóstico, mas esta associação não foi observada em mulheres CT positivas. Infecções por HPV 16 e/ou 18 foram detectadas em 50% das mulheres CT negativas com menos de 29 anos e que apresentavam NIC 2 ou pior diagnóstico, e em 19,5% das mulheres CT negativas com NIC 1 ou não neoplásico. Nestas mulheres, a associação entre HPV 16 e/ou 18 e NIC 2 ou pior diagnóstico foi significativa, mas esta associação não foi observada no grupo CT positivo. Resposta inflamatória e VB foram observadas em 43,5% e 46,2% dos esfregaços cervicais de mulheres com NIC 2. Resposta inflamatória e VB foram observados em 64,2% e 32,6% dos esfregaços cervicais de mulheres com diagnóstico histológico de NIC 3. Nestas mulheres, quando infectadas pelos tipos de HPV 16 e/ou 18, foram observadas resposta inflamatória e VB, respectivamente, em 43,1% e 20% dos casos. Resposta inflamatória apresentou associação estatisticamente significante com NIC 2 ou pior diagnóstico em mulheres infectadas pelos tipos de HPV 16 e/ou 18 (OR= 6,70; 95%IC:2,32-19,31) e por outros tipos de HPV (OR=4,90; 95%IC: 1,86-12,89). Associações significativas foram observadas em mulheres com VB e NIC 2 ou pior diagnóstico, infectadas pelos tipos de HPV 16 e/ou 18 (OR= 3,38; 95% IC :1,07-10,64) e por outros tipos de HPV (OR= 3,38; 95%IC: 1,15-10,01). Conclusões: A infecção por CT detectada por PCR não mostrou associação com o aumento do risco para NIC 2 ou pior diagnóstico em mulheres HPV positivas. Em mulheres CT negativas e com menos de 30 anos de idade, os tipos de HPV 16 e/ou 18 estão associados à NIC 2 ou pior diagnóstico, resultado não observado para as mulheres CT positivas. A VB e resposta inflamatória estão associadas à NIC 2 ou pior diagnóstico em mulheres HR-HPV positivas / Abstract: Objective: To analyze the association between co-infection Human Papillomavirus (HPV) and Chlamydia trachomatis (CT), bacterial vaginosis (BV) and inflammatory response with the severity of the cervical neoplasia. Subjects and methods: This is cross-sectional experimental study carried through in Campinas, São Paulo and in Goiânia, Goiás, Brazil. The casuistic included 290 consecutive women submitted a the Excision of the Transformation Zone or conization due CIN 2 and CIN 3. For the study that evaluated the association between HPV and CT and severity of cervical neoplasia were selected 251 women who were infected with high-risk HPV (HR-HPV). HPV detection .was performed by PCR using primers PGMY09/11 and genotyping by reverse lineblot hybridization assay. The detection of CT was performed by PCR. For the study that evaluated the association between BV and inflammatory response with the severity of cervical neoplasia were selected 211 women infected with HR-HPV with cervical smears available for analysis. The presence of 20% or more clue cells in cervical smears stained by the Papanicolaou method was considered positive for VB. Inflammatory response was assessed by counting the number of neutrophils. The finding of 30 or more neutrophils per field observed under 1000x magnification was taken as presence of inflammatory response. Results: The prevalence of CT in HPV positive women was 15.1% (38/251). Significant association was observed between women with 30 years or older and CIN 2 or worse diagnosis for those CT negative, but this association was not observed for those CT positive. HPV 16 and/or HPV 18 were detected in 50% of the women ? 29 years age with CIN 2 or worse diagnosis who were CT negative, and in 19.5% for those women with CIN 1 or no neoplastic in histological diagnostic. In these women the association between HPV 16 and/or 18 and CIN 2 or worse diagnosis was significative, but this association also was not observed considering the CT positive group. Inflammatory response and BV were observed in 5.5% and 16.7% of cervical smear of women with no neoplastic diagnosis and were observed in 22.9% and 12.5% of cervical smears of women with CIN 1 in histological diagnosis. Inflammatory response and BV were observed in 43.5% and 46.2% of cervical smears of women with CIN 2 in histological diagnosis. The BV prevalence was higher in cervical smears of women infected by the types 16 and/or 18 (25.6%) and inflammatory response was more observed in cervical smears of women infected by other HPV types (25.6%). Inflammatory response and BV were observed in 64.2% and 32.6% of cervical smears of women with CIN 3 in histological diagnosis and were more observed in cervical smears of women infected types 16 and/or 18 representing respectively 43.1% and 20.0% of cases. Inflammatory response and BV were more observed in cervical smears of women infected types 16 and/or 18 in women with invasive carcinoma, representing 27.2% and 18.2% of cases respectively. Inflammatory response was significantly associated with CIN 2 or worse diagnosis in women infeted by HPV16 and/or HPV 18 (OR= 6.70; 95%CI : 2.32-19.31) and HPV other types than HPV16 and/or18 (OR=4.90; 95%CI: 1.86-12.89). Significant associations with BV and CIN 2 or worse diagnosis were observed in women infected by HPV 16 and/or 18 (OR= 3.38; 95%C1:07-10.64) and HPV types other than HPV16 and/or 18 (OR=3.38; 95%CI: 1.15-10.01). Conclusions: CT infection detected by PCR, does not increase the risk for CIN 2 or worse diagnosis in HPV positive women. HPV 16 and/or HPV 18 types in young women are associated with CIN 2 or worse diagnosis, but without obvious association with CT. BV and inflammatory response are associated with CIN 2 or worse diagnosis in women HR-HPV positives women / Doutorado / Ciencias Biomedicas / Doutora em Tocoginecologia

No description available.

Chlamydia trachomatis est une bactérie intracellulaire obligatoire. Ce pathogène de l’Homme est la première cause infectieuse de cécité ainsi que de maladies sexuellement transmissible d’origine bacterienne.Utilisant une souche de C. trachomatis L2 exprimant une protéine fluorescente, nous avons développé des méthodes de microscopie et de cytométrie en flux permettant de suivre les différentes étapes du développement de la bactérie. Ces méthodes faciliteront les futures études de l’infection par Chlamydia.Chlamydia interagit avec différents processus cellulaires, et plus particulièrement via la sécrétion d’effecteurs par le système de sécrétion de type 3 (ST3). Nous avons identifié une famille de protéines possédant un signal de ST3 qui partagent un domaine, le DUF582, présent uniquement chez les Chlamydia pathogènes.Nous avons montré que les 5 protéines DUF582 de C. trachomatis sont exprimées à partir du milieu du cycle infectieux. Nous avons démontré que la protéine Hrs interagit avec le DUF582 et que la protéine DUF582 CT619 interagit avec Tsg101. Hrs et Tsg101 sont d’importants composants de la machinerie ESCRT impliquée dans de nombreux processus de fission membranaire.Utilisant l’interférence ARN, nous avons montré que Hrs et Tsg101 ne sont requis ni pour l’entrée, ni pour le développement de la bactérie. Ceci suggère que les protéines DUF582 bloquent des processus dépendant de Hrs/Tsg101. A l’inverse, la bactérie pourrait utiliser la machinerie ESCRT mais l’existence de mécanismes redondants expliquerait l’absence de phénotype dans les expériences d’interférence. Nous discutons trois hypothèses concernant le rôle des protéines DUF582 dans l’infection. / Chlamydia trachomatis is an obligate intracellular human pathogen. It is the first infectious cause of blindness and the most common cause of sexually transmitted diseases of bacterial origin. Using a strain of C. trachomatis serovar L2 expressing a fluorescent protein we developed microscopy and flow cytometry based methods to quantify several steps of its developmental cycle. These methods will facilitate future studies aimed at testing anti-bacterial compounds or various culture conditions. Chlamydiae interfere with many cellular processes, in particular via the secretion of bacterial proteins through a type 3 secretion (T3S) system. We identified a family of proteins that possess T3S signals. They share a domain designated as DUF582, which is only found in pathogenic chlamydiae. We showed that the five DUF582 proteins of C. trachomatis are expressed from the mid phase of infection. We demonstrated that the protein Hrs is a common interactor for the DUF582. In addition the N-terminal part of the DUF582 protein CT619 interacts with Tsg101. Hrs and Tsg101 are both important components of the ESCRT machinery, which is an ancient machinery required for several processes involving membrane fission.Using RNA interference we showed that Hrs and Tsg101 are dispensable for bacterial entry and growth. This last result suggest that DUF582 proteins actually prevent Hrs and/or Tsg101 driven processes. Alternatively, the bacteria might highjack the ESCRT machinery but redundant mechanisms would explain the absence of phenotype on bacterial development observed in the silencing experiments. We discuss three hypotheses as to the possible role of the DUF582 proteins in infection.

Made available in DSpace on 2015-04-14T13:34:54Z (GMT). No. of bitstreams: 1 422377.pdf: 884424 bytes, checksum: 00e9c69f02bdc012c466f39cf67939e7 (MD5) Previous issue date: 2010-03-04 / A s?ndrome metab?lica (SM) compreende um conjunto de fatores de risco para doen?a ateroscler?tica. Pat?genos como Chlamydia pneumoniae e Helicobacter pylori podem estar envolvidos no deflagramento e perpetua??o da aterog?nese, mas sua rela??o com a SM ? desconhecida. A exposi??o ? Chlamydia trachomatis (CT) em pacientes com SM n?o foi explorada na literatura at? o momento. Objetivo: Avaliar a associa??o, se existente, entre anticorpos IgA e IgG anti-CT e ocorr?ncia de SM. M?todos: Neste estudo transversal, foram avaliados pacientes com SM com e sem eventos card?acos pr?vios e controles sadios pareados por sexo e idade. O diagn?stico de SM se fundamentou nos crit?rios do NCEP. Anticorpos IgA e IgG anti-CT foram detectados por ensaio imunoenzim?tico; t?tulos acima de 1,1 unidades foram considerados positivos para ambos os isotipos. O teste do qui-quadrado foi utilizado para compara??o das vari?veis categ?ricas, e o teste t para compara??o de vari?veis cont?nuas. Para estimar a associa??o entre anticorpos anti-CT e SM, odds ratios (OR) foram calculados. Regress?o log?stica foi utilizada para o ajuste das vari?veis sexo e idade. Resultados: O n?mero total da amostra foi de 238 indiv?duos: 101 pacientes (42,5%) com SM sem eventos card?acos; 47 (19,7%) com SM e eventos card?acos pr?vios; e 90 (37,8%) controles sadios. O sexo feminino predominou nos 3 grupos. A m?dia global de idade foi 59,7 anos. A preval?ncia de teste positivo para IgA anti-CT foi maior em pacientes com SM (17%) do que em controles (6%) (P=0,015). N?veis elevados de IgA anti-CT se associaram significativamente com ocorr?ncia de SM na compara??o com controles sadios ap?s ajuste para sexo e idade (OR=3,4; IC95% 1,2-9,4; P=0,015). Na an?lise de subgrupos, n?veis elevados de IgA anti-CT foram mais prevalentes na SM n?o-complicada do que nos controles (P=0,013). Ap?s ajuste para sexo a idade, a associa??o entre IgA anti-CT e SM n?ocomplicada se manteve definida (OR=3,6; IC95% 1,32-10,2; P=0,015). Conclus?es: Anticorpos IgA anti-CT, possivelmente de fase aguda, foram mais prevalentes na SM do que em controles sadios. Um teste positivo para IgA anti-CT se associou particularmente ? ocorr?ncia de SM n?o-complicada. O papel da resposta IgA anti- CT em pacientes com SM deve ser detalhado em estudos futuros.

Les amibes appartenant au genre Acanthamoeba sont ubiquitaires et responsables de diverses infections, en particulier de kératite amibienne. Par ailleurs elles sont résistantes à de nombreux traitements de désinfection, aussi bien sous leur forme libre (trophozoite) que sous leur forme enkystée. La plupart des données d’efficacité disponibles ont évalué des biocides utilisés pour la désinfection de l’eau potable et/ou des lentilles de contact, mais peu de données sont disponibles concernant le traitement des surfaces ou des matériels médicaux. Ces amibes sont également capables de servir de réservoir à des bactéries pathogènes, notamment des nouvelles espèces de Chlamydia regroupées sous le terme « Chlamydia-like ». Il s’agit de bactéries découvertes récemment et potentiellement responsables d’infections respiratoires et d’avortements spontanés. Malgré leur importance, peu de données sont disponibles concernant la survie et la résistance aux biocides de ces bactéries. L’objectif de ce travail a été dans un premier temps d’évaluer la résistance des amibes aux traitements de désinfections, aussi bien pour les trophozoites que pour les kystes. Dans un second temps, nous avons étudié la survie et la résistance des Chlamydia-like à la désinfection, ainsi que leurs interactions avec les amibes et avec différentes lignées cellulaires ; Chlamydia trachomatis a servi de contrôle dans cette deuxième série d’expérimentations. Les méthodes de culture et d’enkystement des trophozoites ainsi que le choix des souches testées se sont avérés critiques pour l’évaluation des biocides. Certains traitements de désinfection généralement réputés efficaces contre la plupart des micro-organismes ont montré une efficacité limitée vis-à-vis des kystes amibiens ainsi que des trophozoites (glutaraldehyde). Les Chlamydia-like se sont avérées capables de survivre dans l’environnement pendant de longues périodes mais sont globalement sensibles aux désinfectants. Certaines de ces bactéries sont également capables de survivre dans les kystes d’amibes, ce qui peut leur conférer une résistance accrue vis-à-vis des biocides. / Acanthamoebae are ubiquitous amoebae responsible for several infections, mostly amoebic keratitis. They are also resistant to numerous disinfection treatments, as well under their free shape (trophozoite ) as under their encysted shape. Most of the available data of efficiency estimated biocides used for the disinfection of the drinking water and\or the contact lenses, but few data are available concerning the treatment of surfaces or medical devices. These amoebae are also capable of serving as reservoir for pathogenic bacteria, in particular for new species of Chlamydia named " Chlamydia-like ". These new bacteria recently discovered are potentially responsible for respiratory infections and miscarriages. Despite their importance, only few data are available concerning the survival and the resistance to biocides. The objective of this work was at first time to evaluate the resistance of amoebae to disinfection treatments, as well for the trophozoites as for the cysts. Secondly, we studied the survival and the resistance of Chlamydia-like to disinfection, as well as their interactions with amoebae and with various cellular lineages; Chlamydia trachomatis served as control in this second series of experiments. The methods of culture and encystement of trophozoites as well as the choice of selected strains turned out critical for the evaluation of biocides. Some treatments generally considered as effective treatments of disinfection against most of the microorganisms showed an efficiency limited towards the amoebic cysts as well as the trophozoites ( glutaraldehyde ). Chlamydia-like turned out capable of surviving in the environment during long periods but are globally sensitive to disinfectants. Some of these bacteria are also capable to surve in amoebal cysts, what can confer them a resistance increased towards biocides.

The obligate intracellular bacterium, Chlamydia trachomatis, and Herpes Simplex Virus Type-2 (HSV-2) are the leading sexually transmitted pathogens in the world. These infections are usually asymptomatic and clinically mild, but complications can be severe. Reports of dual detection of Chlamydia and HSV within the genital tracts of humans led our laboratory to develop an in vitro Chlamydia/HSV co-infection model. Little is known regarding the specific pathogenesis of Chlamydia and HSV co-infections, but HSV-super-infection of Chlamydia-infected cells caused the chlamydiae to deviate from their normal developmental cycle into a non-replicative state termed persistence, or the chlamydial stress response. Interactions between HSV envelope protein, gD with host cell junction protein, nectin-1, were enough to stimulate the departure from normal chlamydial development. Additional data also suggested that there might be differences between single infection and co-infection outcomes in vivo. Thus, two diverging hypotheses were investigated here: i) that host nectin-1 is required for normal chlamydial development; and ii) that pathogen shedding and/or disease progression in Chlamydia and HSV-2 co-infected animals will differ from that observed in singly-infected animals. Chlamydial infection of nectin-1 knockdown cell lines revealed no inhibition of chlamydial entry, but significant reductions in inclusion size and production of infectious chlamydiae. Additionally, nectin-1 knockout mice shed fewer Chlamydia compared to wild type mice. In other studies, we developed a novel in vivo Chlamydia and HSV-2 intravaginal super-infection model in BALB/c mice. Infection with Chlamydia muridarum, followed up to 9 days later by HSV-2 super-infection, both reduced HSV shedding and protected mice from HSV-induced fatal neurologic disease compared to HSV singly-infected animals. Protection is lost when: i) infected animals are no longer shedding C. muridarum; ii) when mice are inoculated with UV-inactivated C. muridarum; or iii) when viable chlamydiae are eliminated from the genital tract using antibiotics prior to HSV-2 super-infection. Altogether, we have determined that host nectin-1 is required for chlamydial development both in vitro and in vivo, and that chlamydial pre-infection protects mice from subsequent HSV infection. We predict that these observations may lead to novel approaches to prevent human infection by these two common sexually transmitted pathogens.

Background: Chlamydia trachomatis in women is a risk factor for tubal factor infertility and extra uterine pregnancies, but the impact of a C. trachomatis infection on male fertility is unclear. It is also hypothesized that persistent infection with C. trachomatis, or other microorganisms, might initiate/promote ovarian tumor development. The aims of the thesis were to study whether C. trachomatis serum antibodies in women and men had an impact on infertility diagnoses, semen characteristics, pregnancy rates and pregnancy outcomes; furthermore, to explore associations of C. trachomatis, and Mycoplasma genitalium, plasma antibodies with epithelial ovarian cancer and borderline ovarian tumors, as well as the presence of C. trachomatis bacteria, and other microorganisms, in ovarian tissues. Materials and methods: Papers I and II: 244/226 infertile couples were tested for serum C. trachomatis IgG, IgA, IgM and chlamydial Heat Shock Protein 60 (cHSP60) IgG antibodies. C. trachomatis IgG positive couples were also tested for C. trachomatis DNA in a urine sample. The follow-up period was 14-54 months. 244 spontaneously pregnant women were also tested for serum C. trachomatis IgG antibodies. Papers III and IV: Plasma samples from 291 women with epithelial ovarian cancer, borderline ovarian tumors and benign conditions, and plasma samples from 271 healthy controls, were analyzed for C. trachomatis IgG, IgA and cHSP60-1 IgG and M. genitalium IgG antibodies. Ovarian tissues from 186 women with benign ovaries, borderline ovarian tumors and epithelial ovarian cancer, as well as tissues from the contra lateral ovary in 126 women, were analyzed for the presence of C. trachomatis, M. genitalium, Neisseria gonorrhoeae, HPV and the polyoma viruses BKV and JCV with nucleic acid amplification tests. Results: Papers I and II: The prevalence of C. trachomatis IgG antibodies was higher among infertile than fertile women, and there were 9 couples with ongoing C. trachomatis infections. In men, C. trachomatis IgG and IgA antibodies were associated with a reduced likelihood to achieve pregnancy for the couple, as well as lower sperm concentration, reduced sperm motility and vitality, increased teratozoospermia index and the occurrence of leukocytes. C. trachomatis IgG and cHSP60 IgG antibodies in infertile women were associated with tubal factor infertility, but not with reduced pregnancy rates or outcomes. Paper III: cHSP60-1 IgG antibodies were associated with ovarian cancer belonging to the postulated type II pathogenetic pathway when plasma samples obtained more than one year prior to diagnosis were analyzed. M. genitalium IgG antibodies were associated with borderline ovarian tumors; however a statistical type 1 error cannot be excluded. Paper IV: None of the microorganisms studied were found in the ovarian tissue samples. Conclusions: C. trachomatis IgG and IgA antibodies in the man substantially decreases the chances of the infertile couple to achieve pregnancy, and are associated with subtle negative changes in semen characteristics. C. trachomatis IgG and cHSP60 IgG antibodies in the woman are risk factors for tubal factor infertility. Prospective plasma cHSP60-1 IgG antibodies are associated with type II ovarian carcinomas, but C. trachomatis bacteria, or the other microorganisms studied, could not be detected in benign, borderline or malignant ovarian tissues.

Objective: To establish stable transgenic non-toxic plants as a platform for plant-based vaccine production as well as potential oral delivery system of vaccine antigens for sexually transmitted infections (STIs). The concept is to immunize the mucosal immune system present in the gut-associated lymphoid tissues (GALT). HIV-1 p24 subtype C protein has been used as the main antigen model, in parallel with an engineered unique chimeric MOMP antigen from Chlamydia trachomatis serovar E. Methods: Chimeric MOMP and p24 vaccine antigens were successfully inserted into the nuclear genomes of Arabidopsis thaliana and Daucus carota via Agrobacterium-mediated gene transfer. The characteristics of the genetic inserts and corresponding mRNAs and recombinant proteins in planta were described using several methods, including northern, Southern, and western blotting, ELISA, and a commercial HIV Ag/Ab combination assay. Immunogenicity of the antigens was studied in mice models. Results: Transgenes of both plant species expressing p24 or chimeric MOMP were successfully generated. Additional HIV-1 vaccine antigen candidates were introduced and the genetic inserts have been confirmed in Arabidopsis thaliana. The Arabidopsis thaliana expressing p24 and chimeric MOMP were demonstrated to be stable over generations and antigenicity analyses showed that plant-derived HIV-1 p24 and chimeric MOMP retained immunological epitopes when they were expressed in planta. Oral administration of transgenic plant material generated a priming effect of the immune competent cells present in the GALT, shown by the presence of antigen-specific-IgG in mice sera after boosting. Mice immunized with plant-derived HIV-1 p24 antigen were also analyzed for antigen-specific faecal IgA as well as cellular immune responses. However, detectable levels of the two latter immune responses were not observed. The Chlamydia trachomatis chimeric MOMP antigen was further evaluated for its potential as a vaccine antigen candidate, with positive results indicating a more rapid clearance of the Chlamydia trachomatis infection post immunization. Conclusion: Stable non-toxic transgenic plants expressing either HIV-1 p24 or a novel  Chlamydia trachomatis chimeric MOMP antigens have successfully been developed. The two plant-produced STI vaccine antigens have in initial mice feeding studies provided important proof-of-concept for the oral vaccination approach. Now, immunization studies to expand, en-hance, and improve knowledge of the immune responses generated by the orally delivered transgenic plants are of high priority. / Kemi/biokemi

Thesis (Ph. D.)--University of Alabama at Birmingham, 2006. / Title from first page of PDF file (viewed Feb. 14, 2008). Includes bibliographical references.